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Principal Investigator: Ram I. Mahato
Organization: UNIVERSITY OF NEBRASKA MEDICAL CENTER
Fiscal Year: 2024
Award: $390,632
Funding agency: National Institute of Diabetes and Digestive and Kidney Diseases
PROJECT SUMMARY
Liver injury from alcohol abuse, and obesity can lead to liver inflammation, steatosis, and fibrosis. Liver cell
damage results in the release of inflammatory cytokines from hepatocytes and Kupffer cells (KCs), which cause
the activation of hepatic stellate cells (HSCs). If unresolved, it may result in liver fibrosis and progression to
cirrhosis. Hedgehog (Hh) signaling regulates multiple pathways in liver fibrosis, including epithelial-to-
mesenchymal transition (EMT), HSC activation, and inflammation. Further, chronic liver diseases are associated
with the dysregulation of miRNAs. Specifically, miR-96 is upregulated after liver damage and promotes fatty liver
disease (FLD). miR-96 caused malfunctioning of insulin receptor (INSR) and insulin receptor substrate (IRS)-1
results in impaired insulin signaling and glycogen synthesis. Further, miR-96 downregulates SMAD7 and
FOXO1-3 and promotes transforming growth factor-beta 1 (TGF-β1) mediated liver fibrosis. In our preliminary
studies, we demonstrated the upregulation of Hh signaling ligands including PTCH1, SHH, and GLI2 cause
significant increase in collagen and fat deposition in 5% ethanol and high-fat diet (HFD) fed mouse. We
synthesized a novel Hh pathway inhibitor 2-chloro-N 1-[4-chloro-3-(2-pyridinyl) phenyl]-N4, N4-bis(2-
pyridinylmethyl)-1,4-benzenedicarboxamide (MDB5) with GLI1/2 inhibitory activity in nanomolar (nM)
concentration. Treatment of both HFD and alcohol-induced liver disease (ALD) mice with MDB5 resulted in a
significant decrease in the levels of liver injury markers aspartate aminotransferase (AST) and alanine
aminotransferase (ALT), and further ablate GLI2, and its target genes. Our miRNA profiling in ALD and HFD
mouse liver identified miR-96 was consistently upregulated. Target Scan analysis revealed that miR-96 targets
several anti-inflammatory and anti-fibrogenic genes. The knockdown of miR-96 expression in hepatocytes and
HSCs with anti-miR-96 resulted in the restoration of affected genes SMAD7 and FOXO3. We synthesized
glycyrrhetinic acid (GA) conjugated GA-PEG-P(Asp)-g-DC-g-TEPA copolymers for liver-specific in vivo delivery
of MDB5 and anti-miR-96, respectively. There was a significant increase in MDB5 concentration in the fibrotic
liver at 1h post systemic administration of MDB5 loaded GA-NPs. TGF-β and Hh signaling crosstalk whereby
TGF-β1 induces GLI-1 through downstream consequence of RAS signaling. Therefore, we hypothesize that the
combination therapy of MDB5 and anti-miR-96 using GA-NPs could prevent alcohol and fat induced liver injury,
and fibrosis. Our specific aims are to i) establish the therapeutic efficacy of the Hh inhibitor MDB5 on alcohol
and high fat diet induced liver injury; ii) establish the profibrotic role of miR-96 in in HFD and ALD mouse models,
and iii) determine the therapeutic efficacy of liver targeted NPs loaded with MDB5 and anti-miR-96 for treating
HFD and ALD mouse models. Our long-term goal is to understand the progressive mechanisms of ALD and
NAFLD to liver fibrosis, and establish new, liver-specific treatments.
Terms: <ADD-1 protein><ADD1 protein><ALT1><Ablation><Absolute ethanol><Adrenal Cortex Hormones><Adverse effects><Affect><Alanine Aminotransferase><Alanine Transaminase><Alanine-2-Oxoglutarate Aminotransferase><Alcohol Chemical Class><Alcohol abuse><Alcoholic Liver Diseases><Alcoholic steatohepatitis><Alcohols><Animal Model><Animal Models and Related Studies><Anti-Inflammatories><Anti-Inflammatory Agents><Anti-inflammatory><Antioxidants><Apoptosis><Apoptosis Pathway><Aspartate Aminotransferases><Aspartate Apoaminotransferase><Aspartate Transaminase><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Binding><Biodistribution><Bone-Derived Transforming Growth Factor><Causality><Cell Body><Cell Communication and Signaling><Cell Signaling><Cell-Extracellular Matrix><Cells><Cellular injury><Cessation of life><Chemicals><Cirrhosis><Collagen><Combined Modality Therapy><Corticoids><Corticosteroids><DAGAT enzyme><DGAT1><Death><Deposit><Deposition><Development><Diacylglycerol O-acyltransferase 1><Diet><Disease><Disease Progression><Disorder><Drug Kinetics><Drugs><ECM><ETOH><Economic Burden><Enoxolone><Erinaceidae><EtOH abuse><Ethanol><Ethyl Alcohol><Etiology><Evaluation><Extracellular Matrix><FKHR><FKHR-Like 1><FKHRL1><FOXO1><FOXO1A><FOXO1A gene><FOXO3><FOXO3A><FOXO3A gene><Fats><Fatty acid glycerol esters><Fibrosis><Forkhead Box O1A><Forkhead Box O3A><Forkhead in Rhabdomyosarcoma><Forkhead in Rhabdomyosarcoma-Like 1><Formulation><GLI Family Gene><GLI Family Protein><GLI Protein><GLI gene><GLI-1><GLI-Kruppel Family Member 2><GLI1><GLI1 Gene><GLI1 Protein><GLI2><GLI2 gene><Genes><Gli2 protein><Glioma Associated Oncogene Homolog 1 Protein><Glioma Associated Oncogene Homolog Protein><Glioma-Associated Oncogene Homolog><Glutamate-Aspartate Transaminase><Glutamic-Alanine Transaminase><Glutamic-Oxaloacetic Transaminase><Glutamic-Pyruvate Transaminase><Glutamic-Pyruvic Transaminase><Glycogen><Glycyram><Glycyrrhetic Acid><Glycyrrhetinic Acid><Goals><Grain Alcohol><Health><Hedgehogs><Hepatic><Hepatic Cells><Hepatic Disorder><Hepatic Parenchymal Cell><Hepatic Stellate Cell><Hepatocyte><High Fat Diet><IRS-1 protein><Immune reaction><Immunosuppressants><Immunosuppressive Agents><Immunosuppressive drug><Immunosuppressive treatment><Impairment><In Vitro><Inflammation><Inflammatory><Inflammatory Response><Injury to Liver><Insulin Receptor><Insulin Receptor Protein-Tyrosine Kinase><Insulin Receptor Substrate 1><Insulin-Dependent Tyrosine Protein Kinase><Intracellular Communication and Signaling><Ito Cell><Jintan><Kupffer Cells><L-Aspartate-2-Oxoglutarate Aminotransferase><Ligands><Liquid substance><Liver><Liver Cells><Liver Fibrosis><Liver Stem Cell><Liver diseases><MADH7><MADH7 gene><MADH8><Macrophage><Mediating><Medication><Metabolic Diseases><Metabolic Disorder><Methylcarbinol><Mice><Mice Mammals><Micro RNA><MicroRNAs><Milk Growth Factor><Minor><Molecular Interaction><Multimodal Therapy><Multimodal Treatment><Murine><Mus><Myofibroblast><Mφ><NAFLD><NASH><Obesity><Pathogenesis><Pathway interactions><Patients><Pharmaceutical Preparations><Pharmacokinetics><Platelet Transforming Growth Factor><Play><Production><Programmed Cell Death><Regulatory Pathway><Rhetinic Acid><Role><SHH><SHH gene><SMAD7><SREBP-1c><Sampling><Scanning><Signal Induction><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Sonic Hedgehog><Stellate Sinusoidal Macrophage><TGF B><TGF-Beta 1><TGF-Beta1><TGF-beta><TGF-β><TGFB><TGFB1><TGFB1 gene><TGFbeta><TGFβ><Therapeutic><Thesaurismosis><Toxic effect><Toxicities><Transfection><Transforming Growth Factor Beta 1><Transforming Growth Factor beta><Transforming Growth Factor-Beta Family Gene><Transforming Growth Factors><Treatment Efficacy><Tumor Growth Factors><Up-Regulation><Upregulation><Uralenic Acid><Validation><adipogenesis><adiposity><alcohol co-abuse><alcohol induced hepatic injury><alcohol induced liver disorder><alcohol induced liver injury><alcohol prevention><alcohol problem><alcohol related liver disease><alcohol-associated liver disease><alcohol-induced hepatic dysfunction><alcohol-induced liver disease><alcohol-induced liver dysfunction><alcohol-mediated liver dysfunction><alcohol-mediated liver injury><alcohol-related liver disease><alcoholic liver injury><biological signal transduction><causation><cell damage><cell injury><cellular damage><chronic hepatic disease><chronic hepatic disorder><chronic liver disease><chronic liver disorder><cirrhotic><combination therapy><combined modality treatment><combined treatment><copolymer><corpulence><cytokine><damage to cells><developmental><diacylglycerol O-acyltransferase><diacylglycerol acyltransferase><diets><diglyceride acyltransferase><disease causation><drug/agent><effective therapy><effective treatment><epithelial to mesenchymal transition><ethanol abuse><ethanol induced hepatic injury><ethanol induced liver disorder><ethanol induced liver injury><ethanol liver disease><ethanol-induced hepatic dysfunction><ethanol-induced liver disease><ethanol-induced liver dysfunction><ethanol-mediated liver dysfunction><ethanol-mediated liver injury><fatty liver disease><fibrotic liver><fluid><gli2 gene product><glioma associated oncogene 1><glioma associated oncogene family zinc finger 1><glioma associated protein 2><global health><glutamic aspartic transaminase><hazardous alcohol use><hepatic body system><hepatic damage><hepatic disease><hepatic fibrosis><hepatic inflammation><hepatic injury><hepatic organ system><hepatopathy><hepatoprotection><hepatoprotective><immune suppressive agent><immune suppressor><immunoreaction><immunosuppressive substance><immunosuppressor><in vivo><inflamed liver><inhibitor><injury to cells><innovate><innovation><innovative><insulin receptor substrate 1 protein><insulin signaling><intervention efficacy><knock-down><knockdown><lipid biosynthesis><lipogenesis><liquid><liver damage><liver disorder><liver inflammation><liver injury><liver macrophage><liver progenitor><liver repair><mRNA Expression><mRNA seq><mRNA sequencing><mRNA-seq><mRNAseq><metabolism disorder><miRNA><miRNAs><model of animal><mortality><mouse model><multi-modal therapy><multi-modal treatment><murine model><nano formulation><nano medicinal><nano medicine><nano particle><nano-molar><nano-sized particle><nanoformulation><nanomedicinal><nanomedicine><nanomolar><nanoparticle><nanosized particle><non-alcohol fatty liver disease><non-alcohol induced steatohepatitis><non-alcoholic fatty liver disease><non-alcoholic liver disease><non-alcoholic steato-hepatitis><non-alcoholic steatohepatitis><nonalcoholic fatty liver disease><nonalcoholic steato-hepatitis><nonalcoholic steatohepatitis><novel><pathway><pre-clinical evaluation><preclinical evaluation><prevent><prevent alcohol><preventing><preventing alcohol><problem alcohol use><problem drinking><problematic alcohol consumption><problematic alcohol use><repair><repaired><restoration><side effect><social role><sterol regulatory element binding protein-1c><therapeutic efficacy><therapy efficacy><transaminase A><transcription factor ADD1><transforming growth factor beta1><transforming growth factors Animal growth regulators><validations>